A great challenge in microscopic imaging of biological and medical samples (e.g. tissue sections) is to achieve high sensitivity, high specificity, high multiplexing, and high resolution (preferably beyond the diffraction limit) simultaneously. This SBIR proposal addresses this challenge by developing a novel technology platform called Proximity DNA Exchange Imaging. As a result of the studies proposed here, a panel of imaging reagents three high-value cancer-related biomarkers (phosphorylated-AKT, -MET, and -CTNNB1) will be established. Compared to previous and current proximity-based immunoassay platforms, Proximity DNA Exchange Imaging is highly advantageous with regard to ease of implementation, versatility, depth of multiplexed information, and spatial resolution. This technology will offer the research and medical community methods to investigate molecular details of biology with unprecedented precision and information content. It also has broad applications in areas such as digital pathology and high content screening for drug discovery.